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Yeast Smash & Grab DNA Miniprep

based on M.D. Rose, F. Winston, and P. Hieter (1990) Methods in Yeast Genetics: A Laboratory Course Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

1. To cell pellet in Eppendorf tube, add 0.3 g (roughly 0.3 ml) of glass beads, 0.2 ml of lysis buffer and 0.2 ml of a 1:1 mix of phenol and chloroform.
2. Vortex the tube at top speed for 2 min.
3. Add 0.2 ml of TE (10 mM Tris, 1 mM EDTA, pH 8.0) and vortex again for a few seconds.
4. Spin the tubes for 5 min (room temperature) at top speed in an Eppendorf centrifuge.
5. Transfer the aqueous (upper) phase (0.38 ml) to a fresh Eppendorf tube, using a new pipette tip for each sample. Discard the tube with the glass beads.
6. Add 2 volumes of 100% ethanol at room temperature. Mix thoroughly.
7. Centrifuge in Eppendorf for 2-3 min at room temperature.
8. Discard the supernatant (use the aspirator; take care not to dislodge the pellet).
9. Rinse the pellet with 0.5 ml of cold, 70% ethanol add the ethanol slowly down the side of the tube, then centrifuge for 3-5 sec.
10. Remove the supernatant. Leave the tubes open and inverted for the pellets to dry. (Or dry the pellets under vacuum.)


Glass beads

Use 425-600 micron beads. Sigma G-9268 works well for us. To prepare the beads:

Lysis buffer:
10 mM Tris, pH 8.0
100 mM NaCl
1% SDS
2% Triton X-100

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